Journal: PLOS ONE

Article Title: LincROR promotes tumor growth of colorectal cancer through the miR-145/WNT2B/WNT10A/Wnt/β-catenin regulatory axis

doi: 10.1371/journal.pone.0312417

Figure Lengend Snippet: (A), The binding sequence of miR-145 in WNT2B/WNT10A 3’UTR was inserted into the dual-luciferase reporter vector to generate the WT plasmids. In contrast, this binding sequence was deleted to generate MUT plasmids. (B-C), Following transfection with miR-145, the luciferase activities of WT and MUT reporters were assessed. (D-F), The mRNA and protein expression levels of WNT2B and WNT10A were detected in CRC cells with miR-145 or anti-miR-145 transfection into SW620 cells. (G-I) The mRNA and protein expression levels of WNT2B and WNT10A were examined by qRT-PCR and western blot examination in lincROR silencing or overexpressing SW620 cells. Data were shown as mean ± SD (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001; versus the corresponding control group.

Article Snippet: The following antibodies were used to probe membrane at 4°C overnight: β-catenin (1:2000; Cat 8084S, Cell Signalling Technology, USA), WNT2B (1:2000; Cat ab178418, Abcam, UK), WNT10A (1:1000; Cat NBP1-76916, Novus Biologicals, USA), and GAPDH (1:2000; Cat 5174S, Cell Signalling Technology, USA).

Techniques: Binding Assay, Sequencing, Luciferase, Plasmid Preparation, Transfection, Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: PLOS ONE

Article Title: LincROR promotes tumor growth of colorectal cancer through the miR-145/WNT2B/WNT10A/Wnt/β-catenin regulatory axis

doi: 10.1371/journal.pone.0312417

Figure Lengend Snippet: (A-B), the mRNA expression levels of WNT2B and WNT10A were examined in the lincROR overexpressing SW620 cells with miR-145 or anti-miR-145 transfection. (C), The protein expression levels of β-catenin, WNT2B, and WNT10A were examined in the lincROR overexpressing SW620 cells with miR-145 or anti-miR-145 transfection. (D-E), The mRNA expression level of β-catenin was assessed in the lincROR overexpressing SW620 cells with miR-145 or anti-miR-145 transfection. F, The mRNA expression levels of several downstream targets of Wnt/β-catenin signalling were examined in the lincROR overexpressing SW620 cells with miR-145 or anti-miR-145 transfection. Data were shown as mean ± SD (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001; versus the corresponding control group.

Article Snippet: The following antibodies were used to probe membrane at 4°C overnight: β-catenin (1:2000; Cat 8084S, Cell Signalling Technology, USA), WNT2B (1:2000; Cat ab178418, Abcam, UK), WNT10A (1:1000; Cat NBP1-76916, Novus Biologicals, USA), and GAPDH (1:2000; Cat 5174S, Cell Signalling Technology, USA).

Techniques: Expressing, Transfection, Control

Journal: PLOS ONE

Article Title: LincROR promotes tumor growth of colorectal cancer through the miR-145/WNT2B/WNT10A/Wnt/β-catenin regulatory axis

doi: 10.1371/journal.pone.0312417

Figure Lengend Snippet: (A), Representative images of burdened tumors in nude mice (n = 5). (B-C), Tumor weights and volumes were analyzed in the lincROR overexpression group and the control group. (D), The immunofluorescence analysis of Ki-67 was performed on tumor tissues (Scale bar, 20 μm.). (E), The immunohistochemical analysis of β-catenin, WNT2B, and WNT10A was conducted on tumor tissues (Scale bar, 100 μm.). Data were shown as mean ± SD (n = 5). *, P < 0.05; ***, P < 0.001; vs NC; versus the corresponding control group.

Article Snippet: The following antibodies were used to probe membrane at 4°C overnight: β-catenin (1:2000; Cat 8084S, Cell Signalling Technology, USA), WNT2B (1:2000; Cat ab178418, Abcam, UK), WNT10A (1:1000; Cat NBP1-76916, Novus Biologicals, USA), and GAPDH (1:2000; Cat 5174S, Cell Signalling Technology, USA).

Techniques: Over Expression, Control, Immunofluorescence, Immunohistochemical staining

Journal: Nature Communications

Article Title: Autocrine ECM molecules establish MSC quiescence during incisor development by disrupting WNT ligand trafficking process

doi: 10.1038/s41467-025-65705-z

Figure Lengend Snippet: a – d Knockout of Smoc1/2 leads to elevated Wnt activity. a Gene Ontology and KEGG analyses of differentially expressed genes in the mesenchyme of control versus Smoc1/2 DKO incisors at E16.5. b , c Quantitation of expression levels of Axin2 and Lef within individual apical pulp cells of Smoc2 + /- littermate control and Smoc1/2 DKO. Two-tailed Mann–Whitney U test; Axin2 : control vs DKO, p < 0.0001; Lef1 : control vs DKO, p < 0.0001. d Mip-seq analysis showing RNA expression of Axin2 and Lef1 in the incisor of control and Smoc1/2 DKO mice. Scale bars: 50 μm. Representative images from three independent experiments with similar results. e – g Knockout of Smoc1/2 did not significantly affect Wnt6 and Wnt10a RNA levels but changed their protein distribution. e UMAP feature plots showed comparable transcription levels of Wnt6 and Wnt10a in control versus Smoc1/2 DKO incisors at E16.5. f MIP-seq data showed similar RNA expression levels and patterns of Wnt6 and Wnt10a in incisors of Smoc2 + /- littermate controls and Smoc1/2 DKO mice embryos at E16.5. g Antibody staining of WNT6 and WNT10a in incisors of Smoc2 + /- littermate control and Smoc1/2 KO embryos at E16.5. Scale bars: 100 μm. White arrows indicate RNA probe signals in incisor epithelium, while yellow arrows show RNA probe signals in incisors mesenchyme. White asterisks indicate positive immunostaining signals, while yellow asterisks show low signals in the apical pulp regions of incisors. Representative images from three independent experiments with similar results. h , i Quantitation of panels ( f ) revealed no significant difference in the percentage of cells expressing Wnt6 or Wnt10a RNA between control and Smoc1/2 DKO mice incisors. j , k Quantitation of g showed significantly more pulp cells positive for WNT6+ and WNT10a+ immunostaining in Smoc1/2 DKO mice incisors compared to controls. Data are presented as mean ± SEM. n = 3 biological replicates; unpaired two-tailed Student’s t-test. Wnt6 + cell: control vs DKO, p = 0.2071; Wn10a + cell: control vs DKO, p = 0.6739, WNT6+ cell: control vs DKO, p = 0.0003; WNT10a+ cell: control vs DKO, p = 0.0026; N.S.: not significant. l – v Organ culture of incisor tooth germs at E16.5. l The schematic drawing illustrates the experimental scheme for the explant culture. Created with MedPeer (medpeer.cn). Organ culture experiments were conducted using wild-type control incisor explants incubated with WNT6 beads ( m ), WNT6 + SMOC2 beads ( n ), WNT10a beads ( o ), or WNT10a + SMOC2 beads ( p ). The samples were subsequently stained with WNT6 or WNT10a antibodies, respectively, and the staining results were quantified in ( u ). q – t Similar beads explant experiments were performed using Smoc1/2 DKO incisor embryos at E16.5, and the results were quantified in ( v ). Data are presented as mean ± SEM. n = 3 biological replicates; unpaired two-tailed Student’s t -test. WNT6+ cell: control-WNT6 vs control-WNT6-SMOC2, p = 0.4189, control-WNT6 vs DKO-WNT6, p < 0.0001, DKO-WNT6 vs DKO-WNT6-SMOC2, p = 0.0001; WNT10a+ cell: control-WNT10a vs control-WNT10a-SMOC2, p = 0.8690, control-WNT10a vs DKO-WNT10a, p = 0.0003, DKO-WNT10a vs DKO-WNT10a-SMOC2, p = 0.00; N.S.: not significant. Source data are provided as a Source Data file.

Article Snippet: Recombinant Wnt6 (CSB-EP026140MO, CUSABIO) and Wnt10a (CSB-EP026129MO, CUSABIO) proteins were procured for the preparation of Wnt6-beads and Wnt10a-beads.

Techniques: Knock-Out, Activity Assay, Control, Quantitation Assay, Expressing, Two Tailed Test, MANN-WHITNEY, RNA Expression, Staining, Immunostaining, Organ Culture, Incubation

Journal: Nature Communications

Article Title: Autocrine ECM molecules establish MSC quiescence during incisor development by disrupting WNT ligand trafficking process

doi: 10.1038/s41467-025-65705-z

Figure Lengend Snippet: a Expression of active β-Catenin and its downstream targets c-Myc, Cyclin D1, and LEF1 in DPCs after Smoc1 or Smoc2 overexpression, as assessed by western blot. b Expression level of active β-Catenin and its downstream targets c-Myc, and Cyclin D1 in DPCs after Smoc1/2 overexpression alone or simultaneously. c Immunostaining of β-Catenin in control, Wnt agonist WAY and WAY with Smoc1/2 treated groups. Scale bars: 10 μm. Representative images from three independent experiments with similar results. SMOC1/2 interfere with Wnt10a transportation. d The schematic drawing illustrates the experimental scheme of DPCs supplied with beads. Created with MedPeer (medpeer.cn). e Immunostaining of Wnt10a in cultured DPCs supplied with beads. White arrows indicate WNT Dynabeads. Scale bars: 10 μm. Representative images from three independent experiments with similar results. f Expression of active β-Catenin and its downstream targets c-Myc, Cyclin D1, and LEF1 in DPCs after Gpc6 silencing, as assessed by western blot. g Immunostaining of β-Catenin in siRNA negative control (si NC ), WAY with si NC and WAY with si Gpc6 treated groups. Scale bars: 10 μm. Representative images from three independent experiments with similar results. h Co-IP of GPC6 with SMOC1 or SMOC2 in DPCs. i Co-localization analysis of SMOC1/2 with GPC6, as assessed by immunostaining. White arrows indicate co-localization signals. Scale bars: 10 μm. Representative images from three independent experiments with similar results. j Co-IP of GPC6 with WNT10a in DPCs in control , Smoc2 or Smoc1 overexpression groups. Source data are provided as a Source Data file. k Mechanism diagram. SMOC1/2 inhibit WNT trafficking by competitively binding to GPC6, thereby disrupting WNT ligand distribution within the niche. Created with MedPeer (medpeer.cn).

Article Snippet: Recombinant Wnt6 (CSB-EP026140MO, CUSABIO) and Wnt10a (CSB-EP026129MO, CUSABIO) proteins were procured for the preparation of Wnt6-beads and Wnt10a-beads.

Techniques: Expressing, Over Expression, Western Blot, Immunostaining, Control, Cell Culture, Negative Control, Co-Immunoprecipitation Assay, Binding Assay

Journal: Cancer Cell International

Article Title: Identification of the prognostic value of a 2-gene signature of the WNT gene family in UCEC using bioinformatics and real-world data

doi: 10.1186/s12935-021-02215-0

Figure Lengend Snippet: Expression of WNT2 and WNT10A in UCEC. A – B Representativeness of weak WNT2 staining in UCEC (200× and 400×); C – D Representativeness of moderate WNT2 staining in UCEC (200× and 400×); E – F Representativeness of strong WNT2 staining in UCEC (200× and 400×); G – H Representativeness of weak WNT10A staining in UCEC (200× and 400×); I – J Representativeness of moderate WNT10A staining in UCEC (200× and 400×); K – L Representativeness of strong WNT10A staining in UCEC (200× and 400×)

Article Snippet: Rabbit anti-human WNT10A polyclonal antibody was purchased from Proteintech (Wuhan, China).

Techniques: Expressing, Staining

Journal: The FASEB Journal

Article Title: Mapping of Wnt-Frizzled interactions by multiplex CRISPR targeting of receptor gene families

doi: 10.1096/fj.201700144R

Figure Lengend Snippet: Constructs used in this study

Article Snippet: Wnt10a , pcDNA3 Wnt10a , Addgene , 35920 , .

Techniques: Construct, Luciferase

Journal: Respiratory Research

Article Title: Profibrotic role of WNT10A via TGF-β signaling in idiopathic pulmonary fibrosis

doi: 10.1186/s12931-016-0357-0

Figure Lengend Snippet: TGF-β1 induces WNT10A expression in fibroblasts. a The results of an analysis of Hoechst (blue), WNT10A (red) and Phalloidin (green) staining in LL97A cells as indicated by immunofluorescence. The WNT10A expression after the administration of TGF-β1 (5 ng/ml) in LL97A cells was analyzed using a Western blot analysis. b LL97A and IMR-90 cells were treated with TGF-β1 (5 ng/ml), and at the indicated time points, the mRNA level of WNT10A was assessed by real-time qPCR compared to the untreated cells, and the results are expressed as the means ± SEM. * p < 0.05. c The WNT family genes (WNT3A, WNT5A, WNT7B, WNT10A, WNT10B and β-catenin) were analyzed by real-time qPCR compared to the untreated cells, and the results are expressed as the means ± SEM. * p < 0.05. d The luciferase activity of the WNT10A promoter was measured 24 h after TGF-β1 (5 ng/ml) stimulation, and the results are expressed as the means ± SEM. * p < 0.05

Article Snippet: Specific knockdown was achieved using small interfering RNA against WNT10A (sc-76927) or a control siRNA (sc-37007) (all from Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Staining, Immunofluorescence, Western Blot, Luciferase, Activity Assay

Journal: Respiratory Research

Article Title: Profibrotic role of WNT10A via TGF-β signaling in idiopathic pulmonary fibrosis

doi: 10.1186/s12931-016-0357-0

Figure Lengend Snippet: BLM induces WNT10A and β-catenin expressions. a The mRNA level of WNT10A and β-catenin in BLM-treated and control mice was assessed by real-time qPCR ( n = 10 each). * p < 0.05. b Lung tissue specimens of BLM-treated and control mice were stained with anti-WNT10A and β-catenin antibodies. The arrow indicates the WNT10A and β-catenin expressions in fibroblasts. c COS1 cells stably transfected with WNT10A-luciferase reporter plasmid which can replicate in the cells were treated with the indicated concentration of BLM. After 24 h, a luciferase assay was performed and the results are expressed as the means ± SEM. * p < 0.05. d The transfectants described in ( c ) were treated with 50 uM BLM for the indicated times, and then were changed to a new medium. After 48 h, a luciferase assay was performed as a recovery assay, and the results are expressed as the means ± SEM. * p < 0.05

Article Snippet: Specific knockdown was achieved using small interfering RNA against WNT10A (sc-76927) or a control siRNA (sc-37007) (all from Santa Cruz Biotechnology, Inc.).

Techniques: Control, Staining, Stable Transfection, Transfection, Luciferase, Plasmid Preparation, Concentration Assay

Journal: Respiratory Research

Article Title: Profibrotic role of WNT10A via TGF-β signaling in idiopathic pulmonary fibrosis

doi: 10.1186/s12931-016-0357-0

Figure Lengend Snippet: The WNT10A expression regulated collagen production. a Cells were treated with TGF-β1 (5 ng/ml), and at the indicated time points, the mRNA level of COL1A1 was assessed by real-time qPCR. * p < 0.05. b Collagen production was measured 24 h after TGF-β1 (5 ng/ml) stimulation. * p < 0.05. c The efficacy of WNT10A silencing was evaluated by immunoblotting for β-catenin and WNT10A proteins in IMR-90 and LL97A cells. * p < 0.05. d LL97A and IMR-90 cells transfected with indicated siRNAs for 72 h were treated with TGF-β1 (5 ng/ml). After 24 h, the mRNA level of COL1A1 was assessed by real-time qPCR. * p < 0.05. All results are expressed as the means ± SEM

Article Snippet: Specific knockdown was achieved using small interfering RNA against WNT10A (sc-76927) or a control siRNA (sc-37007) (all from Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Western Blot, Transfection

Journal: Respiratory Research

Article Title: Profibrotic role of WNT10A via TGF-β signaling in idiopathic pulmonary fibrosis

doi: 10.1186/s12931-016-0357-0

Figure Lengend Snippet: Comparison between the two groups of IPF patients: positive and negative for WNT10A immunostaining

Article Snippet: Specific knockdown was achieved using small interfering RNA against WNT10A (sc-76927) or a control siRNA (sc-37007) (all from Santa Cruz Biotechnology, Inc.).

Techniques: Comparison

Journal: Respiratory Research

Article Title: Profibrotic role of WNT10A via TGF-β signaling in idiopathic pulmonary fibrosis

doi: 10.1186/s12931-016-0357-0

Figure Lengend Snippet: The clinicopathological significance of WNT10A expression in patients with IPF. a An immunohistochemical analysis of the WNT10A protein expression in the healthy control and patients with IPF. The left panels show normal human lung tissue treated for the localization of WNT10A using immunohistochemistry reactivity in the smooth muscle of vessels (single arrows) and airways (double arrows). The right upper panels show a WNT10A-positive case, which demonstrated a specific expression of WNT10A in one part of the spindled cells (fibroblasts, indicated by arrows in a high-power view of the right panel) at a site of pulmonary fibroblastic foci (boxes, low-power view of left panel). The right bottom panels indicate a WNT10A-negative case in patients with IPF. b The results of the Kaplan-Meier survival analysis of the overall survival in IPF patients who were WNT10A-positive or negative

Article Snippet: Specific knockdown was achieved using small interfering RNA against WNT10A (sc-76927) or a control siRNA (sc-37007) (all from Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Immunohistochemical staining, Control, Immunohistochemistry

Journal: Respiratory Research

Article Title: Profibrotic role of WNT10A via TGF-β signaling in idiopathic pulmonary fibrosis

doi: 10.1186/s12931-016-0357-0

Figure Lengend Snippet: The results of the correlation analysis between WNT10A and the clinical variables in patients with IPF. The Spearman correlation coefficients between these variables were determined as shown on the plots

Article Snippet: Specific knockdown was achieved using small interfering RNA against WNT10A (sc-76927) or a control siRNA (sc-37007) (all from Santa Cruz Biotechnology, Inc.).

Techniques:

Journal: Respiratory Research

Article Title: Profibrotic role of WNT10A via TGF-β signaling in idiopathic pulmonary fibrosis

doi: 10.1186/s12931-016-0357-0

Figure Lengend Snippet: The correlation between WNT10A and the clinical variables in patients with IPF in the WNT10A-positive group. No significant correlations were noted between these clinical variables and positivity of WNT10A based on the Spearman’s correlation coefficients

Article Snippet: Specific knockdown was achieved using small interfering RNA against WNT10A (sc-76927) or a control siRNA (sc-37007) (all from Santa Cruz Biotechnology, Inc.).

Techniques:

Journal: Respiratory Research

Article Title: Profibrotic role of WNT10A via TGF-β signaling in idiopathic pulmonary fibrosis

doi: 10.1186/s12931-016-0357-0

Figure Lengend Snippet: Logistic regression analysis for the factors predicting AE-IPF

Article Snippet: Specific knockdown was achieved using small interfering RNA against WNT10A (sc-76927) or a control siRNA (sc-37007) (all from Santa Cruz Biotechnology, Inc.).

Techniques:

Journal: Respiratory Research

Article Title: Profibrotic role of WNT10A via TGF-β signaling in idiopathic pulmonary fibrosis

doi: 10.1186/s12931-016-0357-0

Figure Lengend Snippet: The role of WNT10A in lung fibrosis. A proposal model depicting the role of WNT10A in lung fibrosis is shown. Injury leads to increased WNT10A expression, either directly or via TGF-β1 signaling, in fibroblasts. Aberrant activation of WNT10A is involved in mediating the fibrogenesis in patients with IPF by inducing the accumulation of collagen and fibronectin

Article Snippet: Specific knockdown was achieved using small interfering RNA against WNT10A (sc-76927) or a control siRNA (sc-37007) (all from Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Activation Assay